Collecting large datasets of rotational electron diffraction with ParallEM and SerialEM

Kiyofumi Takaba; Saori Maki-Yonekura; Koji Yonekura

doi:10.1016/j.jsb.2020.107549

Collecting large datasets of rotational electron diffraction with ParallEM and SerialEM

J Struct Biol. 2020 Aug 1;211(2):107549. doi: 10.1016/j.jsb.2020.107549. Epub 2020 Jun 13.

Authors

Kiyofumi Takaba¹, Saori Maki-Yonekura¹, Koji Yonekura²

Affiliations

¹ Biostructural Mechanism Laboratory, RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.
² Biostructural Mechanism Laboratory, RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan; Advanced Electron Microscope Development Unit, RIKEN-JEOL Collaboration Center, RIKEN Baton Zone Program, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan. Electronic address: yone@spring8.or.jp.

PMID: 32544623
DOI: 10.1016/j.jsb.2020.107549

Abstract

A semi-automated protocol has been developed for rotational data collection of electron diffraction patterns by combined use of SerialEM and ParallEM, where SerialEM is used for positioning of sample crystals and ParallEM for rotational data collection. ParallEM calls standard camera control software through an AutoIt script, which adapts to software operational changes and to new GUI programs guiding other cameras. Development included periodic flashing and pausing of data collection during overnight or day-long recording with a cold field-emission beam. The protocol proved to be efficient and accurate in data collection of large-scale rotational series from two JEOL electron microscopes, a general-purpose JEM-2100 and a high-end CRYO ARM 300. Efficiency resulted from simpler steps and task specialization. It is possible to collect 12-20 rotational series from ~-68° to ~68° at a rotation speed of 1°/s in one hour without human supervision.

Keywords: 3D ED/MicroED; Automated data collection; CRYO ARM; Electron 3D crystallography; Electron diffraction; ParallEM.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Automation
Cryoelectron Microscopy
Data Collection / standards*
Humans
Image Processing, Computer-Assisted / trends*
Microscopy, Electron, Transmission / trends*
Software*